Continuing funds are requested by [7 vision scientists holding 10 NEI R01] grants and their personnel to support [three] resource/service cores. In the [Department of Anatomy/Cell Biology and Ophthalmology]. The cores are central units that enable and enhance vision research at Wayne State University and for [one member, at nearby (15 minutes) Oakland University]. Facilities requested are: Imaging/Histopathology (l/H), [Immunology (I)] and Tissue Culture/Molecular (TC/M). The l/H core will provide for confocal laser scanning [(including 3-D confocal microscopy)], light and electron microscopy, immunofluorescence, and image analysis for Western blots and other cell imaging analyses (e.g., viability, phagocytosis, and quantification of plasma membrane potential), as well as training in equipment use. There also is assistance/training in management of digital images, slit lamp photography and poster and publication production. The I core, in Ophthalmology, will provide sophisticated immunological assays and expert consultation to these vision researchers. It will initiate and stimulate innovative research projects that address emerging questions on the immunological basis of many ocular diseases. Among its functions, it will provide for isolation of blood cells for Staining or viable cryopreservation, immunophenotyping, ELISA evaluation of cytokines/chemokines from tissue lysates and culture supernates, data management and storage, and access to the C6 Accuri Flow cytometer and Cellometer instruments and to the Karmanos Flow facility].The Tissue Culture/Molecular facility will assist and train for preparation of specialized media, isolation, purification and characterization of ocular cells for primary culture, subculture and propagation of established cell lines, cryopreserve cells and [for HEK cell culture for AAV vector packaging and purification, DNA cloning, site directed mutagenesis, and DNA transfection]. Assistance and training Is also available for techniques in molecular biology including PCR array and real time RT-PCR, storage of bacteria, vectors and cDNA constructs. Each core is staffed by a well-trained research assistant and directors are NEI-R01 supported. With the PI, this team functions to enhance NEI-supported studies, and facilitate collaborative efforts in vision.